Preparation method of hair follicle stem cell active factor freeze-dried powder for treating alopecia

A technology for hair follicle stem cells and active factors, which is applied in the field of preparation of lyophilized powder of hair follicle stem cell active factors for the treatment of hair loss, can solve the problems of short storage time, reduced activity, unfavorable transportation, etc. The effect of various activities and easy access to materials

Active Publication Date: 2022-03-01
安徽科门生物科技有限公司
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Problems solved by technology

[0007] The hair follicle stem cell active factor freeze-dried powder is used to replace the hair follicle stem cell factor solution to solve the following problem: some of the cytokines secreted by stem cells will decrease or lose their activity at normal temperature or under the condition of temperature change, and the expected effect cannot be achieved; Stem cell culture medium in liquid state has a short storage time in the greenhouse, which is not conducive to transportation

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  • Preparation method of hair follicle stem cell active factor freeze-dried powder for treating alopecia
  • Preparation method of hair follicle stem cell active factor freeze-dried powder for treating alopecia
  • Preparation method of hair follicle stem cell active factor freeze-dried powder for treating alopecia

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preparation example Construction

[0037] see figure 1 Shown, the preparation method of the hair follicle stem cell active factor freeze-dried powder provided by the invention comprises the following steps:

[0038] S1. Acquisition of hair follicle stem cells: Under sterile conditions, cut the hair root to obtain hair follicle tissue, then wash the hair follicle tissue in DPBS containing double antibody, then cut it into pieces, and inoculate it in the primary medium for primary Subculture, then digest and purify to obtain hair follicle stem cells;

[0039] S2. Acquisition of active factors of hair follicle stem cells: the hair follicle stem cells are subcultured, and then enriched, centrifuged, and concentrated by ultrafiltration to obtain a concentrate of active factors of hair follicle stem cells;

[0040] S3. Mixing and freeze-drying: after mixing the hair follicle stem cell active factor concentrated solution and the auxiliary solution in a mixer, a mixed solution is obtained; after the mixed solution is ...

Embodiment 1-1

[0047] Acquisition of hair follicle stem cells: Under sterile conditions, cut the hair root to obtain hair follicle tissue, then wash the hair follicle tissue in DPBS containing penicillin and streptomycin, and then cut it into pieces (0.8mm 3 ), and inoculated in primary medium for primary culture, then digested and purified to obtain hair follicle stem cells;

[0048] Among them, the primary culture medium is HG-DMEM culture medium containing 10% fetal bovine serum by volume fraction, and the medium is changed every 3 days in the primary culture; the specific operation of digestion is to obtain hair follicles when the cell density is 70-75%. Tissue cells, then add neutral protease, hair follicle tissue cells, and PBS buffer into a centrifuge tube to seal, and place on a shaker at 37°C at 100r / min to digest for 1.5h, centrifuge at 2000rpm, discard the supernatant, and then use sterile PBS buffer Rinse twice with liquid to wash away neutral protease, then add 0.01% trypsin-EDT...

Embodiment 1-2

[0050] Acquisition of hair follicle stem cells: Under sterile conditions, cut the hair root to obtain hair follicle tissue, then wash the hair follicle tissue in DPBS containing penicillin and streptomycin, and then cut it into pieces (0.8mm 3 ), and inoculated in primary medium for primary culture, then digested and purified to obtain hair follicle stem cells;

[0051] Among them, the primary culture medium is HG-DMEM culture medium containing 10% fetal bovine serum by volume fraction, and the medium is changed every 3 days in the primary culture; the specific operation of digestion is to obtain hair follicles when the cell density is 75-85%. Tissue cells, then add neutral protease, hair follicle tissue cells and PBS buffer into a centrifuge tube to seal, and place on a shaker at 37°C at 100r / min for digestion for 2h, centrifuge at 2000rpm, discard the supernatant, and then use sterile PBS buffer Rinse twice to remove neutral protease, then add 0.125% trypsin-EDTA solution, s...

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Abstract

The invention discloses a preparation method of hair follicle stem cell active factor freeze-dried powder for treating alopecia, and belongs to the technical field of cell factors. The preparation method comprises the following steps: S1, obtaining hair follicle stem cells; s2, acquisition of hair follicle stem cell active factors: inoculating hair follicle stem cells into an amplification culture medium for subculture, and then performing enrichment, centrifugation and ultrafiltration concentration to obtain a hair follicle stem cell active factor concentrated solution; s3, mixing and freeze-drying: mixing the hair follicle stem cell active factor concentrated solution with an auxiliary solution in a mixing box to obtain a mixed solution; and carrying out secondary freeze-drying on the mixed solution to obtain the hair follicle stem cell active factor freeze-dried powder for treating alopecia. And the hair follicle stem cell active factor freeze-dried powder is used for replacing a hair follicle stem cell factor solution, so that the problems that a hair follicle stem cell culture solution in a liquid state is short in preservation time in a greenhouse and is not beneficial to transportation are solved.

Description

technical field [0001] The invention belongs to the technical field of cytokines, and in particular relates to a preparation method of freeze-dried powder of hair follicle stem cell active factor for treating alopecia. Background technique [0002] In recent years, affected by various factors such as environment, work pressure, and genetics, the number of people with hair loss has been increasing. Hair loss is due to many internal factors, such as aging or hormonal balance disturbances, as well as various diseases and severe burns or wounds, etc. It is often accompanied by thinning of the skin in the bald area, these symptoms are due to the loss of follicle function. For the treatment of hair follicle-related diseases such as alopecia and alopecia areata, the current conventional treatment methods include gene therapy, drug therapy and hair transplantation. All there is shortcoming for above-mentioned treatment method. Such as drug treatment of alopecia and alopecia areata...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/36A61K9/19A61P17/14C12N5/074C12P21/02
CPCA61K35/36A61K9/19A61P17/14C12N5/0628C12P21/02C12N2509/10C12N2509/00C12N2500/25C12N2501/39C12N2501/11
Inventor 张正亮
Owner 安徽科门生物科技有限公司
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